Abstract
Among the different types of DNA damage that occur endogenously in the cell, depurination is especially prevalent. These lesions can initiate mutagenesis and have been implicated in a variety of diseases, including cancer. Here, we demonstrate a new approach for the detection of depurination at the single-molecule scale using solid-state nanopores. We induce depurination in short duplex DNA using acidic conditions and observe that the presence of apurinic sites results in significantly slower dynamics during electrokinetic translocation. This procedure may be valuable as a diagnostic for in situ quantification of DNA depurination.
Highlights
Depurination is one of the most significant natural mechanisms of DNA degradation, occurring spontaneously under physiological conditions [1]
An estimated 2,000–10,000 purine nucleotides are lost per day in every human cell [2], most often as a result of thermal fluctuations, but potentially through self-catalyzed mechanisms [3] or through the dissociation of DNA adducts [4]
We demonstrate a new assay for the detection of depurination in short duplex DNA using solid-state (SS-) nanopores
Summary
Depurination is one of the most significant natural mechanisms of DNA degradation, occurring spontaneously under physiological conditions [1]. In this process, adenine and guanine bases are liberated when their N-glycosyl linkages to the deoxyribose backbone are hydrolyzed, resulting in an apurinic (AP) site. AP sites have been linked to disease initiation, including cancers [4] and anemias [6]. A technique capable of linking relative DNA damage with various stages of disease could be potentially transformative for diagnosis and treatment of disorders
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